The Establishment Of A Method Of Prescription Screening And Evaluation And Effect Of Nuciferine On 3T3-L1 Preadipocyte Differentiation In Rats

Process of prescriptions is not only the material basis of oral sustained-release formulations, but also it is pivitol points. In vitro accumulation release rate test is the focus of research and development of oral sustained-release formulations, it is also is an important indicator of prescription. Under the premise of the sustained release formulation’s basic technical indicators in line with pharmacopoeia general requirements, to achieve the desired accumulated release rate of formulation and process in vitro is a pre-clinical screening goal, but sometimes accumulated release rate in vitro and drug release behavior in vivo is not consistency. Therefore, screening studies of formulation and process must be combined with animesulideal pharmacokinetic studies, even with clinical trials. Prescription and process should be adjusted according to the results of animal pharmacokinetic studies and clinical trials.Prescription and process which is determined at pre-clinical phase is often the findings of some phase. Verifying the formulation and process through experimental results in vivo is the ultimate target. Accumulated release rate in vitro of oral sustained release formulation is not only an important indicator of prescription and process screening, but also it is an effective and important indicator of product quality control. If the correlation in vivo and in vivo has been established through accumulation release rate test, the results of the accumulated release rate in vitro can predict release behavior in vivo to a certain extent. Therefore, evaluating prescription and process through the necessary preliminary pharmacokinetic study in vivo can decrease technical risks of sustained release formulations to a greater extent.Animal pharmacokinetic studies for the choice of formulation and process is important, its main purpose is to verify whether the agent is of the possibility of burst release or not and evaluate whether release behavior in vivo and in vitro are consistent or not. That is to say, release behavior of drug in vitro is correlation to release behavior of drug in vivo or not.ObjectiveTo establish a method of formulation prescription screening and evaluation. In order to reduce the risk of failure in vitro and to improve the degree of success prescription, we regard the correlation of test in vitro and in vivo as a pivitol indicator of prescription screening. On the one hand, it is useful to screen successful prescription and process, on the other hand, it will be a methological basis of the Beagle dogs for testing and human clinical trials. In this experiment, we take ibuprofen and nimesulide release capsules for examples, we have developed the comparison studies of pharmacokinetics in rat in order to screen optimized prescriptions and process.MethodsFirstly, to establish high performance liquid chromatographic method of oral sustained-release formulations in rats (method specificity, the standard curve and linear regression equation, precision and accuracy test, absolute recovery test, stability test).Secondly, on the basis of preliminary experiments, animesulideals were given oral homemade oral sustained-release formulations and reference preparation (the same active ingredients, specifications and formulations of oral sustained-release formulations). Blood samples of rats were collected at different time and analysized by the appropriate analytical instruments.Thirdly, the relevance of oral homemade oral sustained-release formulations and reference preparation in vivo and in vitro is analysized by the use of DAS software. If the relevance of oral homemade oral sustained-release formulations and reference preparation in vivo and in vitro is not good, prescription revisions will be given to the researchers. ResultsTake ibuprofen sustained release capsules and nimesulide sustained release capsules for example, comparative pharmacokinetics study of oral self-made preparation and reference preparation have been used to verify that consistency of the results in vitro can’t stand for consistency of the results in vivo. Then, we conclude that it is very valuable to screen pharmaceutical prescription and process by the rat model.ConclusionWe established the pharmacokinetic method by cutting rat tail of screening and evaluating sustained release formulation prescription and process which is rarely reported at home and abroad. The method is valuable for selection of formulation prescription and process. It defused the risk of fail Beagle dog and human experiments. Our goal is not to replace the pharmacokinetic study in Beagle dogs and clinical study in patients, but to provide an innovative methodology. It is worth mentioning that it is valuable to evaluate quality of different products which is characteristics of the same active ingredients, formulations and specifications. Currently, obesity is a major hazard to human health problems. As a kind of systemic metabolic diseases, obesity has been paid increasing attention. Obesity has been considered to be a risk factor associated with the genesis or development of various diseases, including cardiovascular disease, type 2 diabetes mellitus and metabolic syndrome, which resulting in an increasing morbidity and mortality. Recent reports have proposed that the preadipocytes play a key role by differentiating into mature adipocytes and increasing fat mass. Obesity is characterized by the accumulation of adipose tissue, which expands due to an increase in adipocyte size and number. Therefore, inhibition of adipogenesis from preadipocytes may regulate the amount of adipose tissue.Nelumbinis Folium, the dried leaf of Nelumbo nucifera Gaertn, is bitter, slightly sweet and neutral. Nelumbinis Folium can be used to relieve summer-heat, to invigorate the spleen function of the spleen and arrest bleeding by reducing heat in blood. It was first recorded in the book of Herbal in Tang Dynasty, the every version of Chinese Pharmacopoeia also has close set. As a traditional chinese medicine, the leaf of Nelumbo nucifera Gaertn is prominent in the diet’s health and treatment of lipid-lowering effect. Studies on antiobesity and lipid-lowering effect of the leaf of Nelumbo nucifera Gaertn have become the focus of attention of scholars in the world. Nuciferine is the main component of the the leaf of Nelumbo nucifera Gaertn extract. However, the literature regarding the effect of alkaloids in the leaf of Nelumbo nucifera for treatment of obesity still remains unclear. In the present study, we investigated the antiobesity potential of Nelumbo nucifera alkaloid (NNA) using 3T3-L1 preadipocytes.ObjectiveThe objective of this study was focused on the induction of apoptosis and the inhibition of preadipocyte proliferation and differentiation by NNA in 3T3-L1 preadipocytes.MethodsWe investigated the effects of Nuciferine on 3T3-L1 preadipocytes in this study. We determined the proliferation of 3T3-L1 preadipocytes by MTT assays and the differentiation of 3T3-L1 preadipocytes by oil red staining, and measured the apoptosis by AnnexinⅤ-FITC/PI staining and Flow Cytometry (FCM).ResultsIn this study we investigated the effects of nuciferine on 3T3-L1 preadipocytes. Results showed that nuciferine significantly inhibited the proliferation and differentiation of 3T3-L1 preadipocytes and its inhibition of proliferation and differentiation in a concentration-dependent and time-dependent manner. At the same time nuciferine also promoted the apoptosis of 3T3-L1 preadipocytes, this was also a concentration-dependent and time-dependent manner.ConclusionWe evaluated the proliferation and differentiation inhibition, apoptosis promotion on 3T3-L1 preadipocytes of nuciferine, and prevention of weight increase in vivo. This is vital important for the further research of nuciferine in vivo and vitro.